Immortalization of swine umbilical vein endothelial cells (SUVECs) with the simian virus 40 large‐T antigen

Implementation of the swine umbilical vein endothelial cells (SUVECs) model in vitro can be instrumental in determining the biology of endothelial cells. We have generated an immortalized endothelial cell line, G‐1410, using Simian virus 40 T‐antigen (SV40 T‐ag) primarily to overcome the short life span before the onset of senescence and high variability among enzymatically isolated cells of primary cultures. Fast proliferating cells were selected from cultures and, after a fifth passage, examined for the presence of the SV40 T‐ag by PCR and immunocytochemistry. Phase contrast and transmission electron microscopy revealed that G‐1410 cells did not differ morphologically from SUVECs. The G‐1410 cells exhibited positive staining for vascular endothelial (VE)‐cadherin and von Willebrand factor (vWF), and formed capillary‐like tube structures on Matrigel. Despite the strong oncogenic signal provided by SV40 T‐ag, these transformed G‐1410 cells have remained karyotypically normal and non‐tumorigenic. G‐1410 cells also responded to stimulation with VEGF, FGF‐2, and newborn calf serum. Moreover, G‐1410 cells showed elevated expression of VEGF120, VEGF164 (VEGF‐A), and FGF‐2 at both mRNA and protein levels. In conclusion, based on the cytological and functional evaluation of the newly obtained immortalized cell line, it can be concluded that G‐1410 cells provide a useful tool for studying the effects of VEGF and FGF systems, and other signal transduction pathways related to angiogenesis. Mol. Reprod. Dev. 78:597–610, 2011. © 2011 Wiley‐Liss, Inc.